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DEVELOPMENT OF IMMUNOHISTOCHEMICAL METHODS FOR THE VALIDATION OF IN VITRO MECHANISTIC EFFECTS OF ENVIRONMENTAL EXPOSURES IN VIVO
Impact/Purpose:
Specifically, this work will support efforts to (1) Identify mechanisms of toxicity for particulate matter (PM) constituents/sources; (2) Describe mechanistic studies of the cellular metabolic and signal pathways in transition metal-induced animal airway injury and inflammation, and relate to human cell findings; and (3) Develop methods for studying mechanisms of effects in tissue from human subjects exposed to PM.
Description:
By necessity, most molecular and biochemical toxicology research is conducted using in vitro models which vary in their ability to approximate toxic responses at the organismal level. Validating the relevance of in vitro mechanistic findings in vivo would lead to increased confidence in the use of mechanistic data. Our laboratory has been developing in vivo methods for the detection of key biochemical and molecular events involved in the toxicity of environmental agents. This work is focused on the use of immunohistochemical staining techniques that use new phosphorylation-state-specific antibodies which recognize only the activated forms of key proteins involved in the transmittal of signals from the outside world to the interior of the cell. To date, we have used this approach to show that the same changes that we have reported to occur in vitro in human airway epithelial cells also occur in the lung tissue of animal exposed to the same metallic pollutants in vivo. Two papers have been published on the development of these methods and their findings. Our ultimate goal is to apply these techniques for the detection of mechanistic endpoints in tissues derived from human subjects undergoing controlled exposures or collected from subjects participating in field studies.